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目的:比较两种抗体纯化方法在分离纯化树鼩IgG抗体的应用,制备抗IgG的多克隆抗体及检测。方法:采用两种商品化IgG抗体纯化试剂盒分离树鼩血清IgG抗体,采用SDS-PAGE和蛋白定量测定提纯IgG。以树鼩IgG作为抗原,与等量弗氏完全佐剂(第一次)、弗氏不完全佐剂(第二次)混合皮下注射免疫兔,对分离血清进行多克隆抗体纯化及Western Blot检测及定量分析。结果:两种方法均能有效分离纯化树鼩IgG,在经过Montage PROSEP-A试剂纯化后的IgG在纯度和含量方面均优于Protein A/G Matrix试剂。通过纯化后的树鼩IgG免疫兔制备的抗IgG抗体能有效识别树鼩IgG。结论:纯化的树鼩IgG具有良好免疫原性,由此制备的抗体具有高度特异性。研究结果为利用树鼩作为实验动物提供了必要的实验基础。
OBJECTIVE: To compare the application of two kinds of antibody purification methods in the isolation and purification of tree gut IgG antibody and to prepare polyclonal anti-IgG antibodies and their detection. Methods: Two commercial IgG antibody purification kits were used to isolate serums from tree shrews. IgG was purified by SDS-PAGE and protein quantification. Rabbit was injected subcutaneously with the same amount of Freund’s complete adjuvant (first time) and Freund’s incomplete adjuvant (second time), and the isolated serum was purified by polyclonal antibody and Western Blot And quantitative analysis. Results: Both methods can effectively isolate and purify tree gut IgG, and IgG purified by Montage PROSEP-A reagent is superior to Protein A / G Matrix reagent in terms of purity and content. Immunization of rabbit with the purified tree gall IgG anti-IgG antibodies can effectively identify tree gall IgG. Conclusion: The purified T.gondiimus IgG has good immunogenicity, and the antibody prepared by it is highly specific. The results provide the necessary experimental basis for using tree trunks as experimental animals.